DE10243837A1 - Process for continuously processing flowable compositions in a flow cell comprises indirectly sonicating the composition in the flow cell via a liquid placed under elevated pressure - Google Patents

Abstract

Continuously processing flowable compositions in a flow cell (4) comprises pre-mixing the components of the flowable composition, and passing and simultaneously sonicating the flowable composition through the cell, and introducing the sonicated composition into a collecting or process vessel. The composition in the flow cell is indirectly sonicated via a liquid (90) placed under elevated pressure. An Independent claim is also included for a flow cell for continuously processing flowable compositions containing a flow path (50) surrounded by a pressure casing (10) connected to an ultrasonic transmitter (60).

Description

The invention relates to a method and a flow cell for the continuous processing of flowable compositions (Working fluid) by means of ultrasound, the flowable composition during the Sound does not come into direct contact with the sounder and from environmental influences Completely can be isolated according to the characteristics of claims 1 and 15.

The invention can be used for continuous, contact- and contamination-free sonication of flowable compositions with Ultrasound can be used.

The method is used, for example in the fields of pharmaceutical technology, food technology, Biotechnology, cell biology, cosmetics, electronics and electrical engineering for the purpose of mixing two or more liquids, for mixing gases and liquids, for emulsifying two immiscible or only partially miscible liquids, for the homogenization of colloidal and coarsely disperse preparations, for disaggregating and crushing solids in a liquid, to control crystal formation (sonocrystallization), for suspension of solids in liquid, for the disaggregation and homogenization of biological organ parts, for cell disruption, for degassing liquids, for control in particular Accelerating chemical reactions, as an intermediate process in the production of liposomes, micellar systems, nano- and microemulsions, Nanoparticles, nanocapsules, microspheres and microcapsules and the like.

Sound waves of frequencies 18 to Approx. 150 kHz (ultrasound) are an energy source that is versatile in process engineering for the manipulation of flowable systems can.

To get away from ultrasound higher frequencies (up to many MHz), which is mainly used for measurement and diagnostic purposes Finding application, delimiting it is also called "power ultrasound".

With sufficiently good transmission the sound energy on the liquid Medium as well as exceeding a system dependent Limit amplitude, cavitation occurs in the sonicated medium. Under Cavitation means the formation of the finest gas bubbles through continuous pressure changes due to the transferred into the fluid Vibration in their size grow and eventually implode. During the implosion, microjets are formed, which too locally concentrated, extremely high pressures and temperatures. The Energy input through cavitation can be used to chemical Initiate reactions, accelerate them or influence their selectivity.

Furthermore, the energy introduced into the liquid system can be used for intensive mixing and for the dispersion of multi-phase systems. It is known to use ultrasound to generate emulsions (e.g. DE 197 56 874 A1 ), for size reduction, deagglomeration and dispersion of solids in liquids (e.g. Vasylliv and Sakka 2001) and for the disintegration of biological material, e.g. cells (e.g. DE 42 41 154 C1 ).

From the DE 197 56 874 A1 A device for producing disperse substance mixtures by means of ultrasound is known, in which the radiation surface of the sonotrode is in direct contact with the liquid to be processed.

From the DE 42 41 154 C1 it is known to disrupt cell dispersions or cell suspensions by means of ultrasound in a flow cell in order to obtain cell contents by the sonotrode projecting into the flow cell by ½ to 2/3 of its length. The immersion angle and the immersion depth of the sonotrode are set depending on the solids content of the medium to be sonicated.

The use is also described of ultrasound to produce liposomes (Arnardóttir et al. 1995). Finally, ultrasound is used to degas liquids Application.

It is widely used Ultrasound in the production of emulsions. In the following, therefore are discussed in more detail in this area of application, but without limit the described invention to this field want. An emulsion is a disperse system of two or several liquids, that do not mix or only partially mix. In the manufacture of Emulsions become common starting from a coarsely disperse raw emulsion which e.g. by means of a simple mixer can be generated, is generally unstable and within broken down into minutes by minutes. By entering more The coarse drops of the raw emulsion become more mechanical energy broken down to the micro or nanometer scale. Because of the engagement of surface active Substances and / or stabilizers can cause coalescence of the droplets and prevents the emulsion from creaming and thus lasts for hours, Be stabilized for months or even years.

It was found that emulsions of comparable quality to those from established processes (high-pressure homogenization) can be produced using ultrasound (Behrend et al. 2000). An increase in energy density (input power per volume times sonication time in batch systems or per volume flow in continuous systems) leads to a reduction in droplet sizes (Bechtel et al. 2000, Behrend et al. 2000). The energy density can be increased either by increasing the power input or by increasing the residence time of the emulsion in the zone of high cavitation intensity.

The discontinuous sound system of liquid and liquid disperse Substance systems are generally made up of the substances in question in a suitable container submitted and if necessary to produce a rough premix touched become. In this liquid a suitable sound generator is then immersed and for a defined one Time activated. Frequently the sounder consists of piezoceramic elements, which by an HF generator can be supplied with an alternating voltage. The AC voltage whose frequency is that of the sound to be generated corresponds to a deformation of the piezoceramics, which thus be put into a mechanical vibration. To the piezoceramic Vibrating elements are coupled to a sonotrode that detects the vibrations possibly reinforced and transferred into the liquid to be sonicated by their end in the liquid is immersed.

A big disadvantage of this procedure is the lack of possibility a scale-up to larger scales. The Space intensely sonicated by the sonotrode is limited roughly on a tapering from the sonotrode surface Cone that only protrudes a few centimeters into the soundproofed room. Liquid, which is outside of this conical space insufficiently influenced by sound. With increasing volume of the vessel to be sonicated becomes the ratio from intensely sonicated room to less influenced room drastically unfavorable.

According to is advantageous DE 197 56 874 A1 the use of a vessel that is only slightly larger in diameter than the radiation area of the sonotrode.

Furthermore, internals or stirring the liquid increase the effectively sonicated volume. Generally beneficial is a continuous arrangement using ultrasound liquid to be charged in a flow cell through the intensely sonicated room directly is forced under the sonotrode. The variation in flow velocity allows an adjustment of the dwell time in the sound zone.

From the patent and scientific A number of such arrangements are known in the literature.

DE 197 56 874 A1 describes, among other things, a flow channel with the diameter of the radiating surface of the sonotrodes used, in which several sonotrodes can be installed in series. Also described there is the connection in series of several flow-through vessels into which sonotrodes protrude.

DE 43 27 679 A1 describes a method for the disruption of cell material by means of ultrasound, in which advantageous conditions such as angles, immersion depths and volume ratios for a sonotrode projecting into a flow cell are mentioned.

Arrangements with flow-through, pot-like vessels of small volume are also known, in which the liquid to be sonicated is guided past a sonotrode ( US 5,032,027 . GB 2 250 930 A. ).

DE 28 46 462 A1 describes a continuous emulsification by means of a flow-through chamber in which a vibrating element is flowed around or sound generators are installed at the edges.

DE 39 30 052 A1 also describes an arrangement in which specially designed reflection walls are arranged opposite the sounders in a flow-through channel.

Common to all these methods is the disadvantage that the sounder is in direct contact with the sonicated medium. The cavitation generated in the sonicated liquid detaches the finest particles from the sonotrode material. After prolonged operation, this is in the form of grooves, holes, etc. Macroscopically visible on the sonotrode surface. The problem is intensified when sonicating suspensions containing solids, since the solid particles have an additional abrasive effect on the sonotrode surface. The described dispensing of particles into the sonicated medium is problematic, for example, when producing products for pharmaceutical purposes. Most commercially available ultrasound sources are equipped with sonotrodes made of metallic alloys. Metallic fine particles and metal ions are thus introduced into the medium exposed to ultrasound. If the product produced by the sonication is intended for medical use, in particular for parenteral administration to humans or animals, or if it is further processed into a product to be administered medically, the metal particles or ions released by the sonotrode represent a safety risk for humans or animals. The same applies to the production of pharmaceuticals that are used on the eye, in the lungs, on areas of skin that have been extensively injured or in body cavities (eg vagina, uterus, bladder). Furthermore, there may be an undesired interaction of the released metal particles and ions with constituents - in particular the active ingredients - of the pharmaceutical product, which in the worst case can thus be inactivated or converted into toxic products. In general, contamination of the sonicated product with substances released from the sonotrode leads to problems if, as explained using the example of the pharmaceutical product, there is an undesirable interaction between the material released from the sonotrode and components of the sonicated material. Undesired interactions would be, for example, hydrolysis, oxidation or reduction, complex formation, aggregation, precipitation, change in the conformation of components of the sonicated material. This can also occur, for example, if cells are used to obtain ingredients (e.g. proteins, Peptides, antibodies, etc.) can be digested. Furthermore, particles and ions released by the sonotrode can have an adverse effect on an analysis of the sonicated product or make it impossible.

Another so far not satisfactory dissolved The problem is an aseptic, i.e. aseptic, problem-free procedure Sound is more fluid Systems. Existing systems either have no hermetic ones Sealing against the environment on and / or are difficult to validate clean and sterilize.

Need for aseptically produced flowable Substance systems exist particularly in the field of pharmaceutical Products, still in cosmetics and food. The latter benefit of an aseptic or at least low-germ production by the addition of preservatives is reduced or avoided entirely can be. The same applies to the manufacture of pharmaceuticals for topical applications, in particular on the injured Skin, on the eye, in the lungs or in body cavities. Pharmaceuticals for parenteral use Administration, which are injected, or pharmaceuticals for use on the injured eye, on large damaged skin areas or for flush of body cavities are imperative be sterile. End sterilization can be disadvantageous or impossible with which an aseptic mode of production is inevitable. Examples for parenteral Administerable pharmaceuticals are infusion solutions for parenteral regulation of the water, electrolyte and carbohydrate balance, preparations for total parenteral nutrition, as well as medicinal preparations (e.g. aqueous or oily solutions, Emulsions, suspensions, microemulsions) and vaccines. Such parenteral preparations are generally administered intravenously, intraarterially, intramuscularly, subcutaneous, intradermal, intraperitoneal, intraocular, intraarticular or intralumbal administered. They often exist from disperse systems or go through in the course of their production one or more dispersion steps. There is in literature Examples using ultrasound to produce such disperse systems was used. When encapsulating hydrophilic active ingredients in biodegradable microspheres for example, one is often used in a first production step Emulsion of aqueous drug solution and organic polymer solution generated, which then become microspheres is processed further. Here is .. the use of ultrasound one By default applied method (e.g. Cohen et al. 1991, Yang et al. 2001). liposomes microspheres and capsules, nanoparticles and capsules and drug-releasing implants are other examples of systems which can go through dispersion steps in their production.

In summary, there is therefore a great potential for an ultrasound system that is able to perform under aseptic conditions disperse material systems without contamination by particles from to produce the sonotrode.

The object of the invention is a Process and a flow cell for the continuous sonication of flowable compositions, especially of small liquid volumes to describe in the flow principle, with which the disadvantages of State of the art can be avoided and with which an efficient Processing a flowable Composition such as mixing, dispersing, emulsifying, suspending, Sonocrystallization, crushing, disagglomeration, cell disruption, extraction, Homogenization, degassing and the like can be ensured without a direct connection between the metallic sonotrode and the one to be sonicated flowable To produce composition, the flowable composition to be sonicated (Working fluid) while the sound system must be kept isolated from environmental influences as required can.

This object is achieved according to the invention the method with the features of claim 1 and with the features the flow cell of claim 15 solved.

The method according to the invention is characterized in that the working fluid in the flow cell via a under elevated Pressurized fluid is sonicated indirectly.

The flow cell according to the invention is characterized in that the flow path through which the working fluid flows is surrounded by a pressure jacket to which an ultrasonic transducer is connected to vibration excitation, where between a outer wall the flow path and the pressure jacket to avoid cavitation one under elevated Liquid under pressure located.

Through indirect vibration excitation a flow path from, for example, a glass or plastic tube (flow cell), in which the flowable composition (working fluid) to be sonicated located about a pressurized liquid in a surrounding pipe, which can also act as cooling, is achieved that no contamination of the flowable composition to be sonicated can be done with metals.

The increased pressure of the liquid prevents cavitation in the liquid and thus transmission losses. The ultrasound acts on the flow cell and with high intensity on the working fluid flowing in it and causes the desired cavitation in the working fluid.

The flow cell is easily replaceable, it can variable, if necessary even smallest volumes in the flow efficiently are processed, there are no anechoic rooms (dead volumes), the flow cell can be easily integrated into existing systems.

The working fluid can be hermetically sealed become.

The system (arrangement) can be in different Dimensions depending realized by the flow rate and the necessary ultrasonic energy become.

The present process is special advantageous because it is the most commonly used ultrasound in many areas flowable Systems without contamination possible. In particular, the invention Do not process the sound energy by direct, as is usual with conventional systems Contact with a vibrating surface with the liquid to be sonicated transferred to this. Instead, the sound energy is, as described above, transferred to a pressurized liquid and passed from there to a flow cell through which the flowable composition to be treated with ultrasound flows.

The described invention is further excellent for sonication of flowable compositions under suitable for aseptic conditions.

The sonicated flowable composition comes exclusively with the pipe in the pressure jacket (flow cell) in contact and is with it during the passage of the ultrasound apparatus hermetically from the environment completed and protected from contamination.

That with the sonicated flowable composition in contact tube (flow cell) can be sterilized very easily become.

Not with the flowable composition Parts of the device in contact can easily be surface disinfected be, for example by spraying / rubbing with disinfectant solutions and / or by treatment with short-wave radiation, e.g. ultraviolet Light.

Assembling the sterilized individual components of the equipment can be unproblematic, for example in a laminar flow work station, isolator or clean room.

Another advantage of the flow cell according to the invention is the possibility that element of the apparatus that has the sonicated flowable composition is in contact, namely the flowed pipe (Flow cell), to be replaced after sonication. A if necessary, the pipe needs to be validated when changing from one product or batch to the next as well a check for possible Wear can thus be avoided. Depending on that for the pipe The material used is this exchange at only a low cost connected that fall below those of cleaning and cleaning validation.

The flow cell according to the invention offers further advantages in that the temperature of the product treated with ultrasound with the aid of a temperature control of the medium flowing through the pressure jacket of the cell can be controlled. Both heating and cooling the sonicated product is possible. Depending on the application temperatures of –80 up to 200 ° C into consideration.

In connection with the implementation of Sonochemical reactions can be used to make the reaction mixture on one for the one to be carried out Reaction favorable Temperature or to dissipate heat generated by an exothermic reaction. You can also use the Temperature influences the properties of the sonicated material become. So can for example, substances melted or in the molten state received, the strength of solids to be divided influenced or thermolabile substances at temperatures below room temperature be sonicated. Also, by dissipating sound energy heat generated in the sonicated material dissipated become.

Appropriate embodiments of the invention are in subclaims described.

The invention is hereinafter in several embodiments the sound of a flowable Composition with ultrasound in a flow cell after the Invention closer explained. In the associated show drawing:

1 : the schematic sectional view of the arrangement for the continuous sonication of a flowable composition (flow cell),

2 : the schematic representation of the process flow using the example of the production of a dispersion of liquids, solids, gases, the raw dispersion being generated in the course of the conveyance into the flow cell and

3 : The schematic representation of the process sequence using the example of the production of a dispersion from liquids, solids, gases by upstream generation of the raw dispersion in front of the flow cell.

The flow cell 4 for the continuous sonication of a small volume of liquid, as shown in the 1 essentially from a tube 10 , in which a preferably non-metallic tube 50 (Flow path, flow cell) with a flowable composition to be sonicated and flowing through 80 (Working fluid) over seals 40 from the pipe 10 beabstan det is arranged.

With the pipe 10 is an ultrasound source for ultrasound excitation 60 connected. With the space between the pipe 10 and the flow cell 50 are a liquid supply 30 and a fluid drain 70 connected by gauges 20 are led.

The pipe 10 is through the ultrasonic transducer 60 vibrated over a liquid 90 , for example water, on the flow cell 50 be transmitted. The liquid 90 is under pressure, for example 4 up to 10 bar to cavitate the liquid 90 and the early wear of the ultrasonic transducer sonotrode 60 and the pipe 10 to prevent. At the same time, the liquid 90 take over the temperature control of the system.

The flow cell 50 vibrates with the liquid 90 and transfers the vibrations to the working fluid in it 80 ,

The final dimensions 20 serve to isolate the seals from vibrations 40 and the connections 30 and 70 for the liquid 90 ,

A change of the flow cell 50 takes place very easily when the device is switched off in the pressureless cooling circuit by releasing clamps (not shown) in the area of the seals 40 ,

To transmit the sound energy from the sonotrode to that with the working fluid 80 flow-through flow cell 50 the following liquids are preferred: water, in particular distilled and deionized, optionally with additives for changing the colligative properties (increase in boiling point, decrease in freezing point); mineral and natural oils and mixtures thereof; Silicone oils and silicone oil mixtures, non-volatile liquids of aromatic or non-aromatic nature and mixtures thereof; Mercury.

As materials for the working fluid to be sonicated 80 flow-through flow cell 50 The following materials are preferred: glass, for example glass type I, II or III according to Europ. Pharmacopoeia, 1997; hard, abrasion-resistant plastics, for example polycarbonate, PVC, polyurethanes, polyamides, polyesters, but also Teflon, polystyrene, polyolefins; ceramic materials, metals and hard metals (if contamination with metal particles and / or ions is not critical).

To transmit the sound energy from the sonotrode to that with the working fluid 80 flow-through flow cell 50 and temperature control of the sonicated working fluid 80 becomes the transmitting fluid 90 according to the invention advantageously set at pressures of 2 to 20 bar, preferably 4 to 10 bar; this at flow rates from 0 to 600 l / h, preferably 0 to 100 l / h and liquid temperatures from -80 to 200 ° C.

The method according to the invention consists in the flowable composition (working fluid) through the ultrasonic flow cell according to the invention 4 to 1 is passed through, sound energy of the ultrasonic flow cell 4 into the working fluid 80 entered and this is changed in the desired manner.

• (1) Dosieren und/oder Vormischen der Komponenten der fliessfähigen Zusammensetzung (Arbeitsflüssigkeit);
• (2) Fördern durch und gleichzeitiges Beschallen der Arbeitsflüssigkeit in der erfindungsgemässen Ultraschalldurchflusszelle;
• (3) Einleiten der beschallten Arbeitsflüssigkeit in ein Auffang- oder Prozessgefäss bzw. Weiterleiten zu einem nachgeschalteten weiteren Verarbeitungsprozess.

The procedure is as shown in the 2 and 3 essentially from three process steps:

• (1) dosing and / or premixing the components of the flowable composition (working fluid);
• (2) conveying by and simultaneously sonicating the working fluid in the ultrasonic flow cell according to the invention;
• (3) Introducing the sonicated working fluid into a collecting or process vessel or forwarding it to a subsequent further processing process.

It is essential to the invention that the working fluid while which three process steps can be isolated from environmental influences and does not come into direct contact with the sonotrode. That's why it can Procedure excluding microbial or non-microbial Contamination and also under strictly defined temperature conditions carried out become.

This has advantages for Treatment more fluid Compositions in the fields of pharmaceutical technology, food technology, Biotechnology, cell biology, cosmetics, electronics and electrical engineering.

The procedure is generally suitable for sonication of one or more immiscible, partially miscible or complete miscible liquids, as well as for the manipulation of one or more liquids and insoluble in it, partially soluble or completely soluble Gases or solids.

The method is particularly suitable for continuous, contact-free and contamination-free sonication of flowable compositions for the purpose of mixing, emulsification, homogenization, comminution, suspension or emulsification of liquids, solids and gases in a conveyable liquid, for cell and organ disruption, for disaggregation, degassing , Implementation and control of chemical reactions, for the control of crystal formation, for the production of liposomes, micellar systems, nano- and microemulsions, nanoparticles, nanocapsules, microspheres and microcapsules and like.

For dosing the components of the flowable Composition (process step 1) are the different components the flowable Composition, for example using suitable pumps, gas cylinders, gas production and gas flow measuring systems, as well as powder conveying and powder dosing systems or also manually into a suitable, preferably hermetically lockable, sterile vessel, Pipe or hose inserted.

The components are then in one preferably externally closed system premixed, which is either in an upstream step before promoting the flowable composition into the flow cell or as part of the funding. A Premixing in the course of the funding the flowable Composition can be done using simple devices such as Tee, suitable mixing valves, frits, membranes, agitated or static Mixer can be achieved.

It is important that the premixing already the components in a sterilizable device or container free of contamination and from environmental influences hermetically shielded.

Funding through and simultaneous Sonication of the flowable Composition in the flow cell according to the invention the (process step 2) is the central step of this invention.

Promoting the flowable composition can be done either before or after premixing the components (process step 1) suitable funding institutions such as pumps or overpressure devices take place, the pumps or overpressure devices must be sterilizable and must work without contamination. The Volume flow of the flowable Composition through the flow cell is controlled so that the flowable Composition during a desired one Time interval in the tube can be sonicated. This time interval Depending on the application, the sound can be between 0.1 seconds and 12 hours, with the preferred sonication time between 1 second and 1 hour. about the vibration amplitude of the sonotrode and its frequency can into the sonicated flowable Composition of registered performance can be checked. Depending on The application of power can be between 1 W and 1 kW can be varied, the power input preferably between 2 and 400 W is. Sound frequencies from 16 to 150 kHz are preferred 20 to 100 kHz, in question.

The dwell time and the performance input determine the flowable composition treated with ultrasound together the registered sound energy, over which together with the Frequency influence on the effect of the sound system can.

For example, it is possible Mean and the width of the droplet size distribution to influence in the production of emulsions.

When crushing solids in a liquid can control the grain size of the solid in the same way become.

There is also the possibility the degree of decomposition when digesting biological material to influence, for example, to separate cell groups into individual cells or to unlock them.

Great importance for the sound effect on the flowable In addition to sonication time, performance input and Sound frequency also the type of material, the geometry and the surface properties the one with the flowable Composition in touch upcoming flow cell as described above under the description of the facility has been.

Can be used as material for this flow cell preferably a non-metal, particularly preferably glass or a hard plastic, chosen to prevent metal particles from entering the sound system flowable To avoid composition.

Furthermore, by suitable choice of materials and / or surface treatment the flow cell undesirable Interactions between components of the flowable composition and the Material of the flow cell can be avoided.

Examples of undesirable interactions are adsorption of components of the flowable Composition to the flow cell, desorption of material components the flow cell (traces of metal, plastic additives), reaction of components of the flowable Composition with the material of the flow cell or desorbed Components of the same, catalytic reactions through the material Flow cell or desorbed flow cell components in the flowable Composition, as well as physico-chemical processes at the interface between the flowable Composition and the flow cell.

By choosing the material for the flow cell or by modifying their surface, the interfacial tension can also between the flow cell and the sonicated flowable composition or components of the same.

Is the flowable composition to be sonicated? of multiple components, so can be the preferred wetting of the flow cell what can be prevented by one or more of these components a mixing of the various components of the flowable composition would be detrimental.

Furthermore, due to the geometry the flow cell or by the nature of the inner surface thereof the flow so changed by the flow cell be that a laminar or turbulent flow of the flowable composition is achieved, which can intensify or weaken the effect of the sound system.

Also has the geometry of the flow cell Influence on the efficiency of sound transmission on the sound system flowable Composition, for example by the wall thickness of the flow cell.

The introduction of the sonicated flowable composition in a collection or process vessel or forwarding to a subsequent further processing process (Process step 3) also takes place according to the invention under precisely controlled ones Conditions and excluding possible contamination.

A single portion can be used as a or multi-serving jar used for intermediate or final storage of the flowable composition become. A process vessel or a forwarding is applied if the flowable composition from the inventive Process is processed in a further process, so like this for example in the production of micro and nanocapsules or micro or nanospheres happens.

The following examples are intended to illustrate the scope of the invention with reference to the schematic representations in FIGS 2 and 3 clarify.

The shows 2 the process sequence using the example of the production of a dispersion from a liquid and optionally a second liquid A, a solid B or a gas C. It becomes a raw dispersion in the course of the conveyance into the ultrasonic flow cell 4 generated.

The 3 illustrates the method according to the invention using the example of the production of an emulsion from a liquid and optionally a second liquid A, a solid B or a gas C. However, the raw dispersion is produced here before being conveyed into the ultrasonic flow cell.

In both figures the continuous liquid phase is included 1 , the dosing of the disperse phase with 2 , a mixer with 3, the supply of the liquid for sound transmission with 30 ( 1 ), a heat exchanger with 6 , the flow of the liquid for sound transmission with 70 ( 1 ), a coarsely dispersive, flowable composition after the process step 1 With 8th and the flowable composition treated by the process (process step 2) 9 designated.

example 1

Preparation of an emulsion of a aqueous solution a model protein in an organic solution of a biodegradable polymer as a preliminary stage for further processing into microspheres as a therapeutic delivery System.

The ultrasound cell 4 comes with a glass tube 50 ( 1 ) with an inner diameter of 2 mm and a wall thickness of 0.5 mm.

The pressure of the liquid 90 in a pressure jacket 10 ( 1 ) the ultrasonic flow cell according to the invention 4 is 4.5 to 5.5 bar, the temperature is approx. 10 ° C.

A 5% (w / w) solution of the polymer poly (milk-co-glycolic acid) in dichloromethane (DCM) and a 10% (w / w) solution of the protein bovine serum albumin (BSA) in a phosphate buffer with pH = 7.4 are used two syringe pumps, not shown, and via a T-piece into the glass tube 50 the ultrasonic cell 4 promoted. The flow rates are 2 ml / h for the BSA solution and 40 ml / h for the polymer solution. The amplitude of the ultrasound cell sounder 4 is varied in the range from 40% to 80% of the maximum amplitude. The W / O emulsions generated are collected and analyzed for their droplet size using laser light scattering (Malvern Mastersizer X, 100 mm lens, Mie diffraction). Reproducibly stable emulsions with an average droplet size (volume distribution) in the range from 1.37 to 0.62 micrometer ( 4 , Table 1), which decreases steadily with increasing amplitude. The emulsions produced at high amplitudes have a very narrow droplet size distribution. The emulsions generated are stable for a period of> 30 minutes and thus fulfill a basic requirement for further processing into microspheres.

Table 1: Average droplet size of emulsions, which were generated as described in Examples 1 to 3 by means of the method according to the invention.

Example 2

The procedure is as in Example 1. In contrast to example 1, the flow rate of the protein solution is 3 ml / h, that of the polymer solution 60 ml / h. The amplitude of the sounder is 80% of the maximum. You get one stable emulsion that opposite Example 1 has only a minimal increase in the average drop size (Table 1) .

Example 3

The procedure is as in Example 1. In contrast to Example 1, dichloromethane is used instead of the solvent becomes the less toxic ethyl formate (EF) as a solvent for the Polymer used. The concentration of the model protein BSA in the buffer solution is 5% (w / w); the flow rates are 2 ml / h for the protein and 40 ml / h for the Polymer solution. The amplitude of the sounder is in the range of 40% to 80% of the Maximum amplitude varies. Reproducibly stable emulsions are obtained with an average droplet size (volume distribution) in the range of 0.90 to 0.68 micrometers (Table 1). With an amplitude a minimum of 60% is found for the mean droplet size. The emulsions have a narrow droplet size distribution and are stable over a period of> 30 minutes.

Example 4

The procedure is as in Example 3. The concentration of the model protein BSA in the buffer solution is 10% (w / w). The polymer and BSA solutions are sterile filtered through a 0.2 micron filter. The glass tube 50 the ultrasonic flow cell 4 , all hose material and the T-piece are autoclaved, the pumps and the ultrasonic flow cell 4 installed in a laminar flow workstation and disinfected by spraying with an ethanolic solution. Sterile disposable syringes are installed in the pumps. The test equipment is assembled in the laminar flow work station. After assembly, the apparatus is rinsed again with an ethanolic solution. It will be like under example 3 described at an amplitude of 80% of the maximum amplitude emulsions. The emulsions produced are diluted with sterile ethyl formate and immediately mixed in a 1:10 ratio with CASO broth and incubated overnight. The following day, the broth is applied to cartridges from the Millipore ^® 100 test system filtered and infested with TSB (tryptic soy broth) medium. According to the European Pharmacopoeia, half of the cartridges are 14 Days at 30-35 ° C to check for bacterial growth, the other half 14 Incubated days at 20-25 ° C to check for fungal growth. No contamination was found.

Example 5

Parenteral oil-in-water (O / W) fat emulsion consisting of soybean oil, an emulsifier, an isotonizing additive and water.

The ultrasonic flow cell according to the invention 4 comes with a sterile glass tube 50 with an inner diameter of 4 mm and a wall thickness of 0.5 mm. The water pressure in the coat 10 the ultrasonic cell 4 is 5 bar, the temperature is 40 ° C. A mixture of 12.0 g of egg lecithin, 22.0 g of glycerin and 866.0 g of water is predispersed in an autoclavable glass bottle with handshakes and then autoclaved according to the 1997 European Pharmacopoeia. In parallel, 100 g soybean oil is also autoclaved in a glass bottle. After cooling, the two liquids are premixed in a mass flow ratio of 10: 1 (water phase oil phase) using peristaltic pumps and sterile tubes in an autoclaved static micromixer and then through the ultrasonic flow cell 4 promoted and sonicated with 80% of the maximum amplitude. The conveying speed is 12 ml / min, which leads to a sonication time of approx. 15 seconds per unit volume of emulsion. The finely dispersed O / W emulsion is filled under aseptic conditions directly into four 250 ml sterile glass bottles for storage and end use as a parenteral fat emulsion. The mean droplet size (volume distribution) of the emulsion produced, determined by means of laser light scattering (Malvern Mastersizer X, 45 mm lens, Mie diffraction), is 405 nm and corresponds to the droplet size of commercially available products.

Example 6

Cell disruption.

The ultrasonic flow cell 4 comes with a sterile glass tube 50 with an inner diameter of 4 mm and a wall thickness of 0.5 mm. The water pressure in the coat 10 the ultrasonic flow cell 4 is 5 bar, the temperature is 4 ° C. A portion of 5 g of sterile, roughly chopped parts of a beef liver is suspended in 100 ml of sterile citrate buffer pH 6.5 and mixed well by vortex. This suspension of cell clumps is passed through the ultrasonic flow cell under aseptic process conditions 4 promoted and sonicated with 90% of the maximum amplitude. The conveying speed is 1.2 ml / min, which leads to a sonication time of approx. 150 seconds per unit volume of cell suspension. The sonicated cell suspension is then transferred to a glass bottle and analyzed. The sonicated cell suspension presents itself as a thick soup, the so-called homogenate. The quality of the digestion is measured on the basis of the glutamate dehydrogenase activity with the substrate 2-oxogluatarate and the activator ADP. A commercially available glutamate dehydrogenase with 120 U / mg enzyme protein is used as a reference. The measured dehydrogenase activity of the cell sample, which was digested according to the method according to the invention, was on average 8-10% above the value obtained after classic digestion by means of detergent or mortar was achieved in a glass flask.

Example 7

liposome Preparation

The ultrasonic flow cell 4 comes with a sterile glass tube 50 with an inner diameter of 2 mm and a wall thickness of 0.5 mm. The water pressure in the jacket of the ultrasonic flow cell 4 is 5 bar, the temperature is 8 ° C. A mixture of 156 mg soy lecithin (0.2 mmol) and 39 mg (0.1 mmol) cholesterol are dissolved in 30 ml dichloromethane methanol (9: 1, v / v) in a 500 ml round bottom flask. The solvent is slowly drawn off in the rotating round-bottom flask, so that a uniform thin lipid film is formed on the flask wall, which is finally dried under vacuum at room temperature. The film is then hydrated with 10 ml of a 100 mM NaCl solution while swirling, producing a cloudy dispersion of large multilamellar vesicles (MLV). The MLV are now conveyed through the ultrasonic flow cell according to the invention and sonicated with 95% of the maximum amplitude. The conveying speed is 1.2 ml / min, which leads to a sonication time of approx. 40 seconds per unit volume of MLV preparation. The sonicated MLV preparation is broken down into small unilamellar lipsomes (so-called SUV, small unilamellar vesicles) by the ultrasound. The liposome preparation is passed through a cellulose acetate membrane filter with a pore size of 0.2 μm installed at the end of the glass tube in order to sterile filter the liposome preparation. Finally, the preparation is made directly in aseptic conditions filled sterile glass vials.

The liposome preparation appears bluish opalescent, which already indicates optically small liposomes. The average liposome size was determined by means of laser light scattering and was 220 nm.

10

pressure shroud (Pipe)

20

final mass

30

liquid inlet

40

seal

50

Flow Cell in the form of a tube

60

ultrasound transducer

70

liquid drain

80

Flowable composition

(Working fluid)

90

Pressure-transmitting and if necessary

thermostatting liquid

1

continuous flowable phase

2

metering the disperse phase

3

mixer

4

Flow Cell

5

-

6

heat exchangers

7

-

8th

coarsely dispersed flowable composition

9

sonicated flowable composition

Claims (18)

A method for the continuous processing of flowable compositions (working fluid) in a flow cell by means of ultrasound, comprising (a) premixing the components of the flowable composition, (b) passing and simultaneously sonicating the flowable composition through the flow cell and (c) introducing the sonicated Working fluid in a collecting or process vessel, characterized in that the working fluid in the flow cell ( 4 ) via a liquid under increased pressure ( 90 ) is sonicated indirectly with ultrasound. A method according to claim 1, characterized in that the working liquid ( 80 ) is thermostatted between - 80 ° C and + 200 ° C during the sonication. A method according to claim 1 and 2, characterized in that the working liquid ( 80 ) is insulated from environmental influences during the sonication and during the phases of premixing and introduction into a collection or process vessel. A method according to claim 1 and 2, characterized in that the working liquid ( 80 ) contains one or more miscible, partially miscible or immiscible liquids and / or one or more soluble, partially soluble or non-soluble gases and / or solids. A method according to claim 1, characterized in that premixing the components of the working fluid in a closed system consisting of a simple T-piece, one Frit, a membrane, a mixing valve or a moving or static mixer, which is tight with the flow cell is connected. A method according to claim 1, characterized in that the working fluid in the flow cell ( 4 ) with a dwell time of 0.1 seconds to 12 hours and the sound energy input between 1 and 1000 W is selected, with a sound frequency of 16 to 150 kHz. Method according to claims 1 to 5, characterized in that a further sonication of the sonicated working fluid ( 80 ) is made. A method according to claim 1, characterized in that the working fluid under aseptic (aseptic) conditions through the flow cell ( 4 ) to be led. Application of the method according to claims 1 to 8 for generating a liquid mixture, for Generating a water-in-oil (W / O) emulsion, oil-in-water (O / W) emulsion, W / O / W or O / W / O double emulsion, a micro or Nanoemulsion, a suspension, a micellar system, or one Lipsomenzubereitung. Application of the method according to claims 1 to 8 for crushing or disaggregating solid particles in a liquid Composition, for the digestion of vegetable or animal Cells for the homogenization and disruption of vegetable and animal tissues, and to break down parasites, bacteria and viruses. Application of the method according to claims 1 to 8 to control crystallization processes (sonocrystallization). Application of the method according to claims 1 to 8 for catalysis or acceleration of chemical reactions. Application of the method according to claims 1 to 8 in processes for the production of microspheres and nanospheres and of micro and nanocapsules. Application of the method according to claims 1 to 8 for degassing or gassing liquid compositions. Flow cell for the continuous processing of flowable compositions (working fluid) by means of ultrasound, containing a flow path, characterized in that the working fluid ( 80 ) flow-through section ( 50 ) from a pressure jacket ( 10 ) is surrounded by an ultrasonic transducer ( 60 ) is connected to the vibration excitation, whereby between an outer wall of the flow path ( 50 ) and the pressure jacket ( 10 ) to avoid cavitation a liquid under increased pressure ( 90 ) flows. Flow cell according to claim 15, characterized in that the flow cell from a tube ( 50 ) for the working fluid ( 80 ) is formed, which is in a of the liquid under increased pressure ( 90 ) flowed tube ( 10 ) via seals ( 40 ) is arranged at a distance from the inner tube wall, which is connected to the ultrasonic transducer ( 60 ) is connected to vibration excitation. Flow cell according to claim 15, characterized in that on the tube ( 10 ) for vibration isolation final masses ( 20 ) are arranged. Flow cell according to claims 15 to 17, characterized in that the liquid ( 90 ) in the tube ( 10 ) is used for temperature control.